M971 chimeric antigen receptors

Abstract

The invention provides a chimeric antigen receptor (CAR) comprising an antigen binding domain comprising SEQ ID NOs: 1-6, a transmembrane domain, and an intracellular T cell signaling domain. Nucleic acids, recombinant expression vectors, host cells, populations of cells, antibodies, or antigen binding portions thereof, and pharmaceutical compositions relating to the CARs are disclosed. Methods of detecting the presence of cancer in a mammal and methods of treating or preventing cancer in a mammal are also disclosed.

1 . An isolated host cell comprising a nucleic acid comprising a nucleotide sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises:
 (a) an antigen binding domain of an antibody which binds to CD22,   (b) a transmembrane domain, and   (c) an intracellular T cell signaling domain,   
       wherein the antigen binding domain comprises an amino acid sequence comprising a heavy chain CDR1 comprising SEQ ID NO: 1, a heavy chain CDR2 comprising SEQ ID NO: 2, a heavy chain CDR3 comprising SEQ ID NO: 3, a light chain CDR1 comprising SEQ ID NO: 4, a light chain CDR2 comprising SEQ ID NO: 5, and a light chain CDR3 comprising SEQ ID NO: 6. 
     
     
         2 . The isolated host cell of  claim 1 , wherein the antigen binding domain comprises a light chain variable region comprising an amino acid sequence comprising SEQ ID NO: 7. 
     
     
         3 . The isolated host cell of  claim 1 , wherein the antigen binding domain comprises a heavy chain variable region comprising an amino acid sequence comprising SEQ ID NO: 8. 
     
     
         4 . The isolated host cell of  claim 1 , wherein the antigen binding domain comprises a linker comprising an amino acid sequence comprising SEQ ID NO: 11. 
     
     
         5 . The isolated host cell of  claim 1 , wherein the antigen binding domain comprises an amino acid sequence comprising SEQ ID NO: 9. 
     
     
         6 . The isolated host cell of  claim 1 , wherein the CAR further comprises a leader sequence. 
     
     
         7 . The isolated host cell of  claim 6 , wherein the leader sequence comprises an amino acid sequence comprising SEQ ID NO: 10. 
     
     
         8 . The isolated host cell of  claim 1 , wherein the transmembrane domain comprises i) a transmembrane portion of a CD8 and/or ii) a transmembrane portion of a CD28. 
     
     
         9 . The isolated host cell of  claim 1 , wherein the transmembrane domain comprises an amino acid sequence comprising SEQ ID NO: 12 or 18 and/or an amino acid sequence comprising SEQ ID NO: 15. 
     
     
         10 . The isolated host cell of  claim 1 , wherein the intracellular T cell signaling domain comprises one or more of i) an intracellular signaling portion of a CD28, ii) an intracellular signaling portion of a CD137, and iii) an intracellular signaling portion of a CD3 zeta. 
     
     
         11 . The isolated host cell of  claim 1 , wherein the intracellular T cell signaling domain comprises an amino acid sequence comprising SEQ ID NO: 16 or 19. 
     
     
         12 . The isolated host cell of  claim 1 , wherein the intracellular T cell signaling domain comprises an amino acid sequence comprising SEQ ID NO: 13 or 20. 
     
     
         13 . The isolated host cell of  claim 1 , wherein the intracellular T cell signaling domain comprises an amino acid sequence comprising SEQ ID NO: 14, 17 or 21. 
     
     
         14 . The isolated host cell of  claim 1 , wherein the CAR comprises an amino acid sequence at least 90% identical to any one of SEQ ID NOs: 22-24. 
     
     
         15 . The isolated host cell of  claim 1 , wherein the intracellular T cell signaling domain comprises an intracellular signaling portion of a CD28 or comprises an intracellular signaling portion of a CD137. 
     
     
         16 . The isolated host cell of  claim 1 , comprising an amino acid sequence at least 90% identical to SEQ ID NO: 22. 
     
     
         17 . The isolated host cell of  claim 1 , comprising and amino acid sequence at least 90% identical to SEQ ID NO: 23. 
     
     
         18 . The isolated host cell of  claim 1 , comprising and amino acid sequence at least 90% identical to SEQ ID NO: 24. 
     
     
         19 . The isolated host cell of  claim 1 , wherein the CAR encoded by the nucleotide sequence is expressed by the isolated host cell. 
     
     
         20 . The isolated host cell of  claim 1 , wherein the isolated host cell is a T cell. 
     
     
         21 . A population of isolated cells comprising at least one isolated host cell of  claim 1 . 
     
     
         22 . A pharmaceutical composition comprising the isolated host cell of  claim 1 , or a population thereof, and a pharmaceutically acceptable carrier. 
     
     
         23 . A method of treating a hematological cancer in a human subject, the method comprising administering to the subject an isolated host cell of  claim 1 , or a population comprising an isolated host cell of  claim 1 , in an amount effective to treat the hematological cancer in the human subject, wherein the isolated host cell is a T cell. 
     
     
         24 . An isolated host cell comprising a nucleic acid comprising a nucleotide sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises:
 (a) a leader sequence comprising an amino acid sequence comprising SEQ ID NO: 10;   (b) an antigen binding domain of an antibody which binds to CD22 comprising an amino acid sequence comprising a heavy chain CDR1 comprising SEQ ID NO: 1, a heavy chain CDR2 comprising SEQ ID NO: 2, a heavy chain CDR3 comprising SEQ ID NO: 3, a light chain CDR1 comprising SEQ ID NO: 4, a light chain CDR2 comprising SEQ ID NO: 5, and a light chain CDR3 comprising SEQ ID NO: 6,   (c) a transmembrane domain comprising an amino acid sequence comprising SEQ ID NO: 12, and   (d) an intracellular T cell signaling domain comprising an amino acid sequence comprising (i) an amino acid sequence comprising SEQ ID NO: 13, (ii) an amino acid sequence comprising SEQ ID NO: 17 or 21, or (iii) both (i) and (ii).   
     
     
         25 . The isolated host cell of  claim 24 , wherein the CAR encoded by the nucleotide sequence is expressed by the isolated host cell. 
     
     
         26 . The isolated host cell of  claim 24 , wherein the isolated host cell is a T cell. 
     
     
         27 . A population of isolated cells comprising at least one isolated host cell of  claim 24 . 
     
     
         28 . A pharmaceutical composition comprising the isolated host cell of  claim 24 , or a population thereof, and a pharmaceutically acceptable carrier. 
     
     
         29 . A method of treating a hematological cancer in a human subject, the method comprising administering to the subject an isolated host cell of  claim 24 , or a population comprising an isolated host cell of  claim 24 , in an amount effective to treat the hematological cancer in the human subject, wherein the isolated host cell is a T cell. 
     
     
         30 . A chimeric antigen receptor (CAR) comprising:
 (a) a leader sequence comprising an amino acid sequence comprising SEQ ID NO: 10;   (b) an antigen binding domain of an antibody which binds to CD22 comprising an amino acid sequence comprising a heavy chain CDR1 comprising SEQ ID NO: 1, a heavy chain CDR2 comprising SEQ ID NO: 2, a heavy chain CDR3 comprising SEQ ID NO: 3, a light chain CDR1 comprising SEQ ID NO: 4, a light chain CDR2 comprising SEQ ID NO: 5, and a light chain CDR3 comprising SEQ ID NO: 6,   (c) a transmembrane domain comprising an amino acid sequence comprising SEQ ID NO: 12, and   (d) an intracellular T cell signaling domain comprising an amino acid sequence comprising (i) an amino acid sequence comprising SEQ ID NO: 13, (ii) an amino acid sequence comprising SEQ ID NO: 17 or 21, or (iii) both (i) and (ii).